Modulation of gene expression in a human bioreactor

ABSTRACT

A method is provided for treating a recipient with a biological product obtained from at least one donor that may be the same as, or different from, the recipient. The method includes identifying a targeted level of gene expression of a first gene in a biological product to be transferred from at least one donor to a recipient; treating the at least one donor to achieve the targeted level of gene expression of the first gene in the biological product; and transferring the biological product from the at least one donor to the recipient.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.16/773,977 filed Jan. 27, 2020, which claims the benefit of U.S.Provisional Application No. 62/797,196, filed Jan. 25, 2019, in the U.S.Patent and Trademark Office, the disclosures of which are incorporatedby reference herein in their entirety.

FIELD OF THE DISCLOSURE

The present disclosure relates generally to gene expression modulation,and more particularly, to systems and methods for inducing targeted geneexpression for a physiological or therapeutic purpose.

BACKGROUND OF THE DISCLOSURE

Transfusion of blood products from a donor to a recipient is a commonmedical procedure, and is frequently employed as an adjunct to surgeryor in the treatment of injuries or diseases. In the past, bloodtransfusions often involved whole blood. At present, however, bloodtransfusions more commonly involve blood components, such as plasma, redblood cells, white blood cells, platelets and clotting factors.

Some recent research efforts have focused on the effect that therelative ages of the donor and recipient have in blood transfusions. Forexample, extensive parabiosis experiments were conducted at theUniversity of California (Berkeley) involving surgically conjoined youngand old mice. Those studies suggested that the blood from a young mousemay rejuvenate tissues in the heart, muscle and brain of the oldermouse. However, it was not clear whether the results of the study werelimited to parabiosis settings.

A subsequent study conducted at Alkahest, Inc. (San Carlos, Calif.)considered the effects of giving blood from young donors (aged 18 to 30)to older subjects (aged 54 to 86) suffering from mild to moderateAlzheimer's disease. The trial involved 18 individuals who were givenweekly infusions for four weeks. Some of the test subjects were given asaline placebo, while others were given plasma from a young donor. Thestudy found no discernible improvements in cognitive abilities in thesubjects treated with young plasma, but did find a significantimprovement in their daily living skills.

SUMMARY OF THE DISCLOSURE

In one aspect, a method is provided for treating a recipient with abiological product obtained from at least one donor. The methodcomprises (a) identifying a targeted level of gene expression of a firstgene in a biological product to be transferred from at least one donorto a recipient; (b) treating the at least one donor to achieve thetargeted level of gene expression of the first gene in the biologicalproduct; and (c) transferring the biological product from the at leastone donor to the recipient. In some variations of this embodiment, therecipient may also be the donor. In such embodiments, the donation maybe autologous, with some time difference between donation and usage ofthe donated sample (for instance by infusion, injection, ortransplantation).

In another aspect, a method is provided for treating a recipient. Themethod comprises (a) obtaining portions of a biological product fromeach of a plurality of donors, wherein each portion of the biologicalproduct is characterized by a level of a first targeted gene expression;(b) determining a desired level of the first targeted gene expression ina derived product to be derived from the obtained portions of thebiological product; (c) forming the derived product by creating amixture of the obtained portions of the biological product having thedesired level of the first targeted gene expression; and (d)administering the derived product to the recipient.

In a further aspect, a method is provided for treating a recipient witha biological product obtained from at least one donor. The methodcomprises (a) determining a targeted level of a first gene expression ina biological product to be transferred from at least one donor to arecipient; (b) selecting at least one donor based on the targeted levelof the first gene expression in the biological product; and (c)transferring the biological product from the at least one donor to therecipient.

In yet another aspect, a method is provided for treating a subjectsuffering from cancer. The method comprises diagnosing the subject assuffering from cancer; upregulating CAMP gene expression in a donor;collecting white blood cells from the donor, thereby obtaining collectedwhite blood cells, wherein the collected white blood cells include Tcells; and performing CAR (Chimeric Antigen Receptor) T cell therapy onthe subject; wherein performing CAR T cell therapy on the subjectincludes (a) genetically modifying the collected white blood cells, and(b) administering the genetically modified white blood cells to thesubject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an illustration of a particular, non-limiting embodiment of aprocess in accordance with the teachings herein.

FIG. 2 is an illustration of another particular, non-limiting embodimentof a process in accordance with the teachings herein.

DETAILED DESCRIPTION

It has now been found that beneficial effects may be realized indonor/recipient therapies involving the donation of a biological productby modulating one or more gene expressions in the donor prior todonation of the biological product. Without wishing to be bound bytheory, it is believed that this approach may be utilized to achievemore optimal levels of one or more therapeutic agents in the biologicalproduct (and/or the body or tissues of the recipient) than may bepossible without gene expression modulation in the donor.

While this approach may be especially suitable to donors of blood orblood components, it is not limited thereto, and may be applicable todonors of many other biological products. Such biological products mayinclude, without limitation, red blood cells, white blood cells,plateless, plasma, clotting factors, cells, organs or tissues.

In a preferred embodiment, a method is provided for treating a recipientwith a biological product obtained from at least one donor. The method101, which is summarized in FIG. 1 , preferably includes identifying atargeted level of a first gene expression in a biological product to betransferred from at least one donor to a recipient 103. This targetedlevel of the first gene expression may be determined, for example, fromresearch, or by performing tests (such as, for example, blood tests orassays) on an intended recipient. The at least one donor is then treatedwith a suitable treatment to achieve the targeted level of the firstgene expression in the biological product 105, after which thebiological product is transferred from the at least one donor to therecipient 107.

The targeted level of gene expression may consider various factors. Forexample, in applications in which the biological product is bloodplasma, the targeted level of the first gene expression may be based onlevels of the first gene expression which will increase the shelf lifeof peptides or enzymes in the blood plasma.

Various gene expressions and gene expression products may be utilized inthe systems and methodologies disclosed herein. These include, withoutlimitation, hormones (including gender-specific hormones such astestosterone and estrogen), antioxidants, protease inhibitors, proteins,metabolites, fatty acids, prostaglandin, alkaloids, peptides, LL-37 andits protein precursor hCAP-18, Proteinase 3, Sirtuin-1, NAD+, BDNF(Brain Derived Neurotrophic Factor), PGC-1a, (Anti-Oxidant ResponseElement), CoQ10, and GHK.

It will be appreciated that the systems and methodologies disclosedherein are not limited to those involving a single donor. Thus, forexample, in some embodiments, one or more pools of donors may beprovided. In such embodiments, a biological product may be derived fromcontributions by individual members of the one or more pools. Forexample, such a derived biological product may be obtained by blendingcontributions from individual members of the one or more pools in such amanner as to obtain a derived biological product that has targetedlevels of one or more gene expressions. In some embodiments, thisapproach may be utilized to achieve a gene expression profile orfingerprint consisting of a plurality of gene expressions G_(e)∈[G_(el),. . . , G_(en)], wherein each G_(i)∈G_(e) is a targeted gene expressionthat may correspond, for example, to a desired numerical range for thegene expression.

It will also be appreciated that the systems and methodologies disclosedherein contemplate the possibility that the donor and recipient may bethe same person. For example, in some embodiments, the donation may beautologous. In such embodiments, there may be a time interval betweendonation and usage of the donated sample, and usage of the donatedsample may involve, for example, infusion, injection, ortransplantation.

In the systems and methodologies described herein, the donor may betreated in various ways to achieve a targeted level of gene expression.For example, the donor may be given a pharmaceutical composition whichmay include, for example, resveratrol, niacin, niacinamide, amino acids(such as, for example, 5-HTP (5-Hydroxytryptophan), also known asoxitriptan), mucuna, omega-3 fatty acids (such as, for example, DHA(docosaliexaenoic acid)), alphalipoic acid, sulforaphane, SOD-I, GPx,catalyse, thymosin, butyrate, phenylbutyrate, Vitamin D3, retinoids thatmay serve as Retinoid X Receptor Agonists, curcuminoids, and variouscombinations or subcombinations of the foregoing. The donor may alsoundergo suitable treatment or stimuli to induce the formation, in thebody of the donor, one or more of the foregoing materials. The donor mayalso be requested to adhere to a pharmaceutical regiment (such as, forexample, taking prescribed pills or capsules over a prescribed timeinterval), which may involve administration of any of the materialsdisclosed above; to adopt a prescribed diet, exercise regiment orlifestyle; or to undergo exposure to predetermined stimuli such as, forexample, electromagnetic or emotional stimuli.

With respect to the use of emotional stimuli, it is to be noted thatdirect emotions stimuli are the result of the sensorial stimulusprocessing by cognitive mechanisms. When an event occurs in anenvironment and is witnessed by a subject, sensorial stimuli arereceived by the subject. The cognitive mechanisms process this stimulusand generate the emotional stimulus for each one of the emotions to beaffected. Examples of emotional stimuli include, but are not limited to,emotional words, emotional images, emotional faces, and emotional filmclips. See, e.g., Grühn, Daniel & Sharifian, Neika (2016), “Lists ofEmotional Stimuli”, which is incorporated herein by reference.

In some embodiments, the donor may be subjected to genome editing (alsoknown as gene editing) or RNA modification. In such embodiments, DNA maybe inserted, deleted, modified or replaced in the genome of the donor.Preferably, such editing will involve targeting genomic modifications tosite specific locations.

This may involve, for example, the use of engineered nucleases, or“molecular scissors”, to create site-specific double-strand breaks(DSBs) at desired locations in the genome. The induced double-strandbreaks may then be repaired through nonhomologous end-joining (NHEJ) orhomologous recombination (HR), resulting in targeted mutations or“edits”. The engineered nucleases used for this process may include, butare not limited to, meganucleases, zinc finger nucleases (ZFNs),transcription activator-like effector-based nucleases (TALEN), and theclustered regularly interspaced short palindromic repeats (CRISPR/Cas9)system. Examples of possible RNA modification pathways include thosedescribed, for example, in Machnicka M A, Milanowska K, Osman Oglou O,et al. MODOMICS: a database of RNA modification pathways—2013 update.Nucleic Acids Res. 2013; 41 (Database issue):D262-D267, which isincorporated herein by reference in its entirety.

The donor (or donors) in the systems and methodologies described hereinmay be selected in accordance with various criteria. These criteria mayinclude, for example, the stipulation that the donor (or donors) are ofthe same or different sex than the recipient; that the difference in age(ΔA) between the donor(s) and recipient is less than, or greater than,some threshold value; that a gender-specific hormone (which may include,for example, estrogen or testosterone) is absent in or is present in (orpresent at some specified or minimum level or range in) the donor(s); onthe basis of a genomic analysis (which may involve, for example,ascertaining the presence of genomic features in both the donor and therecipient, or may involve choosing a donor based at least partially onthe presence of genomic features in the donor that are missing in therecipient); on the presence (or absence) of gene expressions in thedonor(s); or on the basis of the metabolics (or relative metabolics) ofthe donor and recipient. For example, in some applications, it may bedesirable to match donors and recipients more closely in age such that,for example, ΔA≤5, ΔA≤3 or ΔA≤1. In other applications, it may bedesirable for there to be a significant difference in age between donorsand recipients (especially if, for example, the recipient is an elderlyperson) such that, for example, ΔA»10, ΔA≥15 or ΔA≥20.

The pharmaceutical compositions utilized in the systems andmethodologies disclosed herein may utilize one or more activeingredients (and will preferably utilize multiple active ingredients)which may be dissolved, suspended or disposed in various media. Suchmedia may include, for example, various liquid, solid or multistatemedia such as, for example, emulsions, gels or creams. Such media mayinclude liquid media, which may be hydrophobic or may comprise one ormore triglycerides or oils. Such media may include, but is not limitedto, vegetable oils, fish oils, animal fats, hydrogenated vegetable oils,partially hydrogenated vegetable oils, synthetic triglycerides, modifiedtriglycerides, fractionated triglycerides, and mixtures thereof.Triglycerides used in these pharmaceutical compositions may includethose selected from the group consisting of almond oil; babassu oil;borage oil; blackcurrant seed oil; black seed oil; canola oil; castoroil; coconut oil; corn oil; cottonseed oil; evening primrose oil;grapeseed oil; groundnut oil; mustard seed oil; olive oil; palm oil;palm kernel oil; peanut oil; rapeseed oil; safflower oil; sesame oil;shark liver oil; soybean oil; sunflower oil; hydrogenated castor oil;hydrogenated coconut oil; hydrogenated palm oil; hydrogenated soybeanoil; hydrogenated vegetable oil; hydrogenated cottonseed and castor oil;partially hydrogenated soybean oil; soy oil; glycerol tricaproate;glycerol tricaprylate; glycerol tricaprate; glycerol triundecanoate;glycerol trilaurate; glycerol trioleate; glycerol trilinoleate; glyceroltrilinolenate; glycerol tricaprylate/caprate; glyceroltricaprylate/caprate/laurate; glycerol tricaprylate/caprate/linoleate;glycerol tricaprylate/caprate/stearate; saturated polyglycolizedglycerides; linoleic glycerides; caprylic/capric glycerides; modifiedtriglycerides; fractionated triglycerides; and mixtures thereof. The useof coconut oil is especially preferred.

Various fatty acids may be utilized in the pharmaceutical compositionsdisclosed herein. These include, without limitation, both long and shortchain fatty acids. Examples of such fatty acids include, but are notlimited to, docosahexaenoic acid, caprylic acid, capric acid, lauricacid, butyric acid, and pharmaceutically acceptable salts thereof.

The pharmaceutical compositions disclosed herein may be applied invarious manners. Thus, for example, these compositions may be applied asoral, transdermal, transmucosal, intravenous or injected treatments, orvia cell-based drug delivery systems. Moreover, these compositions maybe applied in a single dose, multi-dose or controlled release fashion.

The pharmaceutical compositions disclosed herein may be manufactured astablets, liquids, gels, foams, ointments or powders. In someembodiments, these compositions may be applied as microparticles ornanoparticles.

In some embodiments, the pharmaceutically acceptable compositionsdisclosed herein preferably include a mixture of at least four morepreferably at least five, and most preferably at least six materials(preferably active materials) selected from the group consisting ofphenylbutyrate, bexarotene, curcumin, resveratrol, retinol,cholecalciferol, fatty acids, and pharmaceutically acceptable saltsthereof. In other embodiments, the pharmaceutically acceptablecompositions disclosed herein preferably include a mixture of at leastfour more preferably at least five, and most preferably at least sixmaterials (preferably active materials) selected from the groupconsisting of phenylbutyrate, bexarotene, curcumin, resveratrol,retinol, phenylbutyrate, cholecalciferol, docosahexaenoic acid, caprylicacid, capric acid, lauric acid, and pharmaceutically acceptable saltsthereof.

Various counterions may be utilized in forming pharmaceuticallyacceptable salts of the materials disclosed herein. One skilled in theart will appreciate that the specific choice of counterion may bedictated by various considerations. However, the use of sodium andhydrochloride salts may be preferred in some applications.

While the systems, methodologies and compositions disclosed herein maybe utilized to treat human subjects, it will be appreciated that thesesystems, methodologies and compositions may also be utilized to treatnon-human subjects. In particular, it is contemplated that thesesystems, methodologies and compositions may find use in treating animalsubjects in veterinary applications, or in research or laboratorysettings.

The systems and methodologies disclosed herein have variousapplications. For example, Findlay et al., “Exposure to theantimicrobial peptide LL-37 produces dendritic cells optimized forimmunotherapy”, Oncolmmunology (2019), shows that dendritic cellsexposed to LL-37 become potentiated to be ideal for infusion to enhancethe effectiveness of CAR T cell therapy. This is an important approachfor cancer treatment that is currently in development. However, in theapproach of Findlay et al., the dendritic cells were treated exogenousto the body. In accordance some embodiments of the methodologiesdisclosed herein, the CAMP gene may be upregulated within the body of adonor (which may be the same as, or different from the patient), whichwould affect the phenotype of the “donor's” endogenous dendritic cells.These cells may then be utilized for treatment.

More specifically, in embodiments of the methodologies disclosed herein,CAMP gene expression may be upregulated in a donor. White blood cells(including T cells) may be collected from that donor, and these whiteblood cells may be genetically transformed or modified through CAR(Chimeric Antigen Receptor) T cell therapy (this therapy typicallyinvolves the addition of the gene for a special receptor that binds to acertain protein on the patient's cancer cells) to cause the T cells torecognize or be receptive to particular markers or cellular ligands thatmay be present on cancer cells. Such markers may include, for example,PD-1 or PD-L1 (PD-1 is a protein found on T cells that regulates thebody's immune responses in that, when it is bound to PD-L1 (anotherprotein), it helps keep T cells from killing other cells, includingcancer cells). The subject may then be treated using the methodologiesdisclosed herein for upregulating CAMP gene expression, after which themodified CAR T cells may be re-infused into the subject (here, it isnoted that upregulated endogenous LL-37 expression will preferably haveactivated the subject's dendritic cells in such a way that CAR T celltherapy may work better). In other words, in this particular embodiment,if the donor is the subject, then the subject is treated to unregulatedCAMP gene both before and after the steps of removing T cells and thenre-infusing transgenic CAR T cells.

The conventional CAR T cell therapy accomplishes two objectives. Firstof all, the T cells are genetically altered so that they are receptiveto the particular markers (e.g., PD-1 or PD-L1) found on cancer cells.Secondly, the T cells are propagated in culture so there is a greaternumber of them. Hence, the success of the therapy is premised on the Tcells being focused on killing cancer cells in a subject, and on therebeing more of them. The paper by Findlay et al. shows that T cellspropagate more efficiently when exposed to LL-37 (although, aspreviously noted, in the approach of Findlay et al., the dendritic cellswere treated exogenous to the body).

However, it has now been discovered that LL-37 induction may be utilizedin a donor prior to CAR T cell therapy so that, when the resulting Tcells are collected from the donor, they are better primed to propagateefficiently. It has further been discovered that LL-37 induction may beutilized in a recipient prior to the point when the expanded(propagated) T cells generated from the CAR T cell therapy areadministered to or infused into the body of the recipient. This secondLL-37 induction serves to activate the dendritic cells, which activatethe cytotoxic T cells.

Upregulation of LL-37 typically lasts, at most, about 24 hours. In atypical CAR T cell therapy, there is a 2-4 week interval between thetime when the T cells are harvested, and the time when the geneticallytransformed T cells are returned or reinfused to the subject. With thefirst LL-37 induction, it is preferred that the white blood cells areharvested within 12 hours of when the first LL-37 induction occurs (thatis, within 12 hours of the time at which a pharmaceutical composition isadministered to the subject which upregulates LL-37).

It is preferred that the second LL-37 induction occurs within 24 hoursbefore, and more preferably within 10 hours before, the time when the Tcells are returned or reinfused to the subject. More preferably, thesecond LL-37 induction occurs within 6 hours, even more preferablywithin 4 hours, and most preferably within 3 hours, before the time whenthe T cells are returned or reinfused to the subject.

The CART cell therapy may target the CD19 antigen. It may be utilized,within the context of the systems and methodologies disclosed herein, totreat various cancers including, without limitation, relapsed/refractoryB-cell precursor acute lymphoblastic leukemia (ALL) andrelapsed/refractory diffuse large B-cell lymphoma (DLBCL).

The systems and methodologies disclosed herein may effectively utilizeany of the generations of CARs developed to date. Thus, in someembodiments, the CAR may include an extracellular binding domain, ahinge region, a transmembrane domain, and at least one intracellularsignaling domain. In other embodiments, the CAR may include at least oneco-stimulatory domain, which may be selected, for example, from thegroup consisting of CD28, 4-1BB, CD28-41BB and CD28-OX40. In still otherembodiments, the CAR may include at least one cytokine selected from thegroup consisting of IL-2, IL-5 and IL-12.

The foregoing methodology is summarized in FIG. 2 . As seen therein, apreferred embodiment of the methodology 201 includes diagnosing asubject as having cancer 203. Then, CAMP gene expression is upregulatedin a donor 205 (which may be the same as, or different from, thesubject) using the methodologies disclosed herein. White blood cells arethen collected 207 from the donor and are utilized in performing CAT Tcell therapy on the subject 209, which will typically include (a)genetically modifying the collected white blood cells, and (b)administering the genetically modified white blood cells to the subject.

Another application of the systems, methodologies and compositionsdisclosed herein relates to the treatment of a recipient with abiological product (such as blood, blood plasma or cells) obtained froma donor. In some embodiments, the donor and recipient may be the same.In other embodiments, the donor may include a plurality of donors. Inthis application, the CAMP gene is upregulated in the donor(s) beforethe biological product is collected. The collected biological productmay then be infused into the recipient. In some embodiments, there maybe a significant age difference between the recipient and the donor(s).

Another application of the systems, methodologies and compositionsdisclosed herein relates to the treatment of recipients havingdegenerative conditions. Such degenerative conditions may include, butare not limited to, Alzheimer's disease, amyotrophic lateral sclerosis(ALS, Lou Gehrig's disease), cancers, CharcotMarie Tooth disease (CMT),chronic traumatic encephalopathy, cystic fibrosis, certain cytochrome coxidase deficiencies (these may be the cause, for example, ofdegenerative Leigh syndrome), degenerative disc disease (DDD),EhlersDanlos syndrome, fibrodysplasia ossificans progressive,Friedreich's ataxia, frontotemporal dementia (FTD), certaincardiovascular diseases (for example, atherosclerotic diseases such ascoronary artery disease or aortic stenosis), Huntington's disease,infantile neuroaxonal dystrophy, keratoconus (KC), keratoglobus,leukodystrophies, macular degeneration (AMD), Marfan's syndrome (MFS),certain mitochondrial myopathies, mitochondrial DNA depletion syndrome,multiple sclerosis (MS), multiple system atrophy, muscular dystrophies(MD), neuronal ceroid lipofuscinosis, NiemannPick diseases,osteoarthritis, osteoporosis, Parkinson's disease, pulmonary arterialhypertension, prion diseases (such as, for example, Creutzfeldt-Jakobdisease or fatal familial insomnia), progressive supranuclear palsy,retinitis pigmentosa (RP), rheumatoid arthritis, psoriatic arthritis,plaque psoriasis, lupus nephritis, lupus erythematosus, SandhoffDisease, spinal muscular atrophy (SMA, motor neuron disease), subacutesclerosing panencephalitis, TaySachs disease, and various forms ofdegenerative dementia.

A specific example of the use of the systems, methodologies andcompositions disclosed herein in treating a degenerative condition(Alzheimer's disease) may be appreciated with respect to commonlyassigned U.S. Ser. No. 16/038,158, now published as U.S. 2019/0015361(Barron et al.), “POLYTHERAPY MODULATING CATHELICIDIN GENE EXPRESSIONMODULATION FOR THE TREATMENT OF ALZHEIMER'S DISEASE AND OTHERCONDITIONS”, which is incorporated herein by reference in its entirety.There, a polytherapy of orally available compounds is disclosed thatsynergistically modulates and induces the expression of the cathelicidingene (CAMP), which encodes the host defense peptide LL-37. By providinga number of different CAMP-inducing compounds together at the same time,stronger gene induction is achieved than with just one or two compounds,because the mechanism of induction broadens. The approach of the '158application may be applied in accordance with the teachings herein byhaving a donor (or donors) of a biological product (for example, bloodor blood plasma) undergo the polytherapy described in the '158application prior to donation to induce the expression of the LL-37peptide. The polytherapy will preferably be administered within 10 hoursof the donation, more preferably within 3-6 hours of the donation, andmost preferably within 3-4 hours of the donation. In some cases, therecipient may also undergo the polytherapy, preferably within the sameor similar time frame as the donor(s). This approach may ensure elevatedlevels of the LL-37 peptide in the body of the recipient post-infusion.

One skilled in the art will appreciate that the gene expression (andtargeted level of that gene expression) that will be appropriate intreating a degenerative condition will depend on the particular diseaseor condition being treated. By way of example, although the precisepathophysiology of DDD is not fully understood, the progressive declinein aggrecan (the primary proteoglycan of the nucleus pulposus) appearsto be implicated in the disease. Consequently, imbalance in thesynthesis and catabolism of certain critical extracellular matrixcomponents may be mitigated by the transfer of genes to intervertebraldisc cells encoding factors that modulate synthesis and catabolism ofthese components.

Another application of the systems, methodologies and compositionsdisclosed herein relates to the treatment of recipients having toxigenicconditions. Examples of toxigenic conditions include, but are notlimited to, diphtheria, anthrax infections and Clostridium difficileinfections (CDI). These conditions may involve various pathogens suchas, for example, C. ulcerans, A. flavus, B. anthracis, Corynebacteriumand C. difficile.

Another application of the systems, methodologies and compositionsdisclosed herein relates to the treatment of recipients havingpathogenic conditions. Examples of pathogenic conditions include, butare not limited to, ocular neurodegenerative diseases (such as, forexample, AMD, glaucoma/ocular hypertension, and diabetic retinopathy),including those involving the pathogenic tau protein.

The destructive effects of the pathogenic form of tau can be exerted byseveral forms of tau such as soluble oligomers, insoluble aggregates, oreven by cis p-tau which is an early driver of pathogenic tau. Hence,targeted therapy based on pathogenic tau could be a promisingtherapeutic for patients with CNS or ocular neurodegeneration, to notonly prevent the more destructive effects of pathogenic tau, but alsorestore normal functioning to neural cells.

The above description of the present invention is illustrative, and isnot intended to be limiting. It will thus be appreciated that variousadditions, substitutions and modifications may be made to the abovedescribed embodiments without departing from the scope of the presentinvention. Accordingly, the scope of the present invention should beconstrued in reference to the appended claims.

101. A method for treating a recipient, comprising: obtaining portionsof a biological product from each of a plurality of donors, wherein eachportion of the biological product is characterized by a level of a firsttargeted gene expression; determining a desired level of the firsttargeted gene expression in a derived product to be derived from theobtained portions of the biological product; forming the derived productby creating a mixture of the obtained portions of the biological producthaving the desired level of the first targeted gene expression; andadministering the derived product to the recipient.